肺炎支原体肺炎患儿外周血CXCL8及其mRNA表达
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摘要
目的:探讨支原体肺炎患儿外周血CXCL8及其mRNA表达的临床意义。方法:收集2013年10月~2015年3月淮南市妇幼保健院收治的支原体肺炎患儿48例,其中重症12例,轻症36例,以ELISA法检测患儿血清CXCL8含量,PCR法检测患儿外周血单个核细胞内CXCL8 mRNA水平。以GAPDH为参照,以lgc DNA/lg GAPDH比值代表其最终mRNA水平。结果:支原体肺炎患儿外周血血清CXCL8含量及外周血单个核细胞内CXCL8 mRNA水平分别为(298.917±51.860)pg/ml、(1.848±0.525)lgc DNA/lg GAPDH,与正常对照相比差异均有显著统计学意义(P<0.05)。进一步观察发现,重症患儿外周血CXCL8及其mRNA进一步升高,与轻症组相比,血清CXCL8含量差异无显著统计学意义统计学意义(P>0.05),而CXCL8mRNA水平差异有显著统计学意义(P<0.05)。急性期以红霉素静脉注射7~10 d,使患儿病情得以明显控制,咳嗽症状减轻,肺部炎症逐渐改善,病情得到有效控制,再以阿奇霉素序贯治疗2~3周,患儿病情逐步由急性期转为恢复期,此时患儿外周血CXCL8及其mRNA水平明显降低,与急性期相比,差异有显著统计学意义(P<0.05)。结论:支原体肺炎患儿外周血CXCL8及其mRNA表达水平增高,并与病情的严重程度相关。CXCL8参与支原体肺炎的发病过程,并对病情的轻重程度和转归有一定的提示作用。阿奇霉素可通过抑制肺炎支原体增殖途径降低患儿血清中CXCL8含量、下调CXCL8 mRNA的表达,逐渐抑制由肺炎支原体介导的免疫损伤。
Objective: To study the expression of CXCL8 in the serum and CXCL8 mRNA in the peripheral blood mononuclear cells( PBMCs) of the children with Mycoplasma pneumoniae pneumonia( MPP) and its clinical significance. Methods: Forty-eight children( severe cases 12,light cases 36) with MPP were recruited from October 2013 to March 2015 in the Maternal and Child HealthCare Hospital of Huainan. The concentration of the CXCL8 in serum and the level of CXCL8 mRNA in the PBMCs were measured by enzyme linked immunosorbent assay( ELISA) and polymerase chain reaction( PCR). Taking GAPDH as the internal reference,the ratio of lgc DNA / lg GAPDH was regarded as the extreme level of CXCL8 mRNA. Results: The serum level of CXCL8 and expression of CXCL8 mRNA in PBMCs in the children with MPP were( 298. 917 ± 51. 860) pg / ml and( 1. 848 ± 0. 525) lgc DNA / lg GAPDH. Compared with the normal control,there were significant differences between the two groups( P < 0. 05). Further observation showed that the levels of CXCL8 in serum were no significant difference between in light cases and severe illness( P > 0. 05). However,the expression of CXCL8 mRNA in peripheral blood of the children with severe illness was significantly higher than those in light cases( P < 0. 05). Intravenous infusion of Erythromycin was provided in the acute phase for seven to ten days,so that the children' s condition could be significantly controlled,and the symptoms of pulmonary inflammation were also relieved. Followed by the use of sequential therapy of Azithromycin for about two to three weeks,the children's condition were gradually from acute stage to recovery stage. At this time,the CXCL8 and its mRNA levels in peripheral blood of the sick children were all significantly decreased comparing with those in the acute stage( P < 0. 05). Conclusion: The expression of CXCL8 and its mRNA were increased in the peripheral blood of the sick children with Mycoplasma pneumonia,and also correlated with the severity of the disease. CXCL8 can participate in the pathogenesis of Mycoplasma pneumonia,and has a certain cue effect on the severity and prognosis of the disease. Azithromycin can reduce the content of CXCL8 in serum of the sick children via the pathway of inhibiting the proliferation of Mycoplasma pneumoniae,and down regulate the expression of mRNA,so that the immune injury mediated by Mycoplasma pneumoniae may be gradually inhibited.
Objective: To study the expression of CXCL8 in the serum and CXCL8 mRNA in the peripheral blood mononuclear cells( PBMCs) of the children with Mycoplasma pneumoniae pneumonia( MPP) and its clinical significance. Methods: Forty-eight children( severe cases 12,light cases 36) with MPP were recruited from October 2013 to March 2015 in the Maternal and Child HealthCare Hospital of Huainan. The concentration of the CXCL8 in serum and the level of CXCL8 mRNA in the PBMCs were measured by enzyme linked immunosorbent assay( ELISA) and polymerase chain reaction( PCR). Taking GAPDH as the internal reference,the ratio of lgc DNA / lg GAPDH was regarded as the extreme level of CXCL8 mRNA. Results: The serum level of CXCL8 and expression of CXCL8 mRNA in PBMCs in the children with MPP were( 298. 917 ± 51. 860) pg / ml and( 1. 848 ± 0. 525) lgc DNA / lg GAPDH. Compared with the normal control,there were significant differences between the two groups( P < 0. 05). Further observation showed that the levels of CXCL8 in serum were no significant difference between in light cases and severe illness( P > 0. 05). However,the expression of CXCL8 mRNA in peripheral blood of the children with severe illness was significantly higher than those in light cases( P < 0. 05). Intravenous infusion of Erythromycin was provided in the acute phase for seven to ten days,so that the children' s condition could be significantly controlled,and the symptoms of pulmonary inflammation were also relieved. Followed by the use of sequential therapy of Azithromycin for about two to three weeks,the children's condition were gradually from acute stage to recovery stage. At this time,the CXCL8 and its mRNA levels in peripheral blood of the sick children were all significantly decreased comparing with those in the acute stage( P < 0. 05). Conclusion: The expression of CXCL8 and its mRNA were increased in the peripheral blood of the sick children with Mycoplasma pneumonia,and also correlated with the severity of the disease. CXCL8 can participate in the pathogenesis of Mycoplasma pneumonia,and has a certain cue effect on the severity and prognosis of the disease. Azithromycin can reduce the content of CXCL8 in serum of the sick children via the pathway of inhibiting the proliferation of Mycoplasma pneumoniae,and down regulate the expression of mRNA,so that the immune injury mediated by Mycoplasma pneumoniae may be gradually inhibited.
引文
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[11]Kobayashi Y,Wada H,Rossios C,et al.A novel macrolide solithromycin exerts superior anti-inflammatory effect via NF-κB inhibition[J].J Pharmacol Exp Ther,2013,345(1):76-84.
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[14]毕慧娟,王健,黄河胜,等.慢性乙肝患者外周血单个核细胞中CXCR1、CXCR2及IL-8 mRNA表达与干扰素治疗关系[J].细胞与分子免疫学杂志,2012,28(4):422-425.
[15]Jornot L,Cordey S,Caruso A,et al.T lymphocytes promote the antiviral and inflammatory responses of airway epithelial cells[J].PLo S One,2011,6(10):e26293.
[16]Liu L,Gao Z,Xia C,et al.Comparative study of trans-oral and trans-tracheal intratracheal instillations in a murine model of acute lung injury[J].Anat Rec(Hoboken),2012,295(9):1513-1519.
[17]Jundi K,Greene CM.Transcription of interleukin-8:how altered regulation can affect cystic fibrosis lung disease[J].Biomolecules,2015,5(3):1386-1398.
[18]Gottipati KR,Bandari SK,Nonnenmann MW,et al.Transcriptional mechanisms and protein kinase signaling mediate organic dust induction of IL-8 expression in lung epithelial and THP-1 cells[J].Am J Physiol Lung Cell Mol Physiol,2015,308(1):L11-L21.
[19]Wang J,Lu Q.Expression of T subsets and m IL-2R in peripheral blood of newborns with hypoxic ischemic encephalopathy[J].World J Pediatr,2008,4(2):140-144.
[20]Tsivkovskii R,Sabet M,Tarazi Z,et al.Levofloxacin reduces inflammatory cytokine levels in human bronchial epithelia cells:implications for aerosol MP-376(levofloxacin solution for inhalation)treatment of chronic pulmonary infections[J].FEMS Immunol Med Microbiol,2011,61(2):141-146.
[21]Curatolo W,Foulds G,Labadie R.Mechanistic study of the azithromycin dosage-form-dependent food effect[J].Pharm Res,2010,27(7):1361-1366.
[22]Tanaka R,Sanada S,Fujimura M,et al.Ventilatory impairment detection based on distribution of respiratory-induced changes in pixel values in dynamic chest radiography:a feasibility study[J].Int J Comput Assist Radiol Surg,2011,6(1):103-110.
[23]舒林华,尚云晓,蔡栩栩,等.肺炎支原体肺炎患儿支气管肺泡灌洗液和血清中肺表面活性蛋白的变化及意义[J].中国当代儿科杂志,2012,14(12):928-932.
[24]马香,丁明杰,赵秀侠,等.不同胸部影像学表现的肺炎支原体肺炎儿童的肺功能变化特征[J].中国当代儿科杂志,2014,16(10):997-1000.
[2]Sun H,Chen Z,Yan Y,et al.Epidemiology and clinical profiles of Mycoplasma pneumoniae infection in hospitalized infants younger than one year[J].Respir Med,2015,109(6):751-757.
[3]Lin CH,Nai PL,Bien MY,et al.Thrombin-induced CCAAT/enhancer-binding proteinβactivation and IL-8/CXCL8 expression via MEKK1,ERK,and p90 ribosomal S6 kinase 1 in lung epithelial cells[J].J Immunol,2014,192(1):338-348.
[4]中华医学会儿科学分会呼吸学组.儿童社区获得性肺炎管理指南(2013修订)(上)[J].中华儿科杂志,2013,51(10):745-752.
[5]中华医学会儿科学分会呼吸学组.儿童社区获得性肺炎管理指南(2013修订)(下)[J].中华儿科杂志,2013,51(11):856-862.
[6]王健,韩忠燕,周娜.CXCL8及其受体CXCR1、CXCR2在慢性乙肝患者外周血PMNs中的表达[J].中国免疫学杂志,2015,31(3):375-379,383
[7]Hausner M,Schamberger A,Naumann W,et al.Development of protective anti-Mycoplasma pneumoniae antibodies after immunization of guinea pigs with the combination of a P1-P30 chimeric recombinant protein and chitosan[J].Microb Pathog,2013,64:23-32.
[8]Chang HY,Jordan JL,Krause DC.Domain analysis of protein P30in Mycoplasma pneumoniae cytadherence and gliding motility[J].J Bacteriol,2011,193(7):1726-1733.
[9]Romero-García J,Francisco C,Biarnés X,et al.Structure-function features of a Mycoplasma glycolipid synthase derived from structural data integration,molecular simulations,and mutational analysis[J].PLo S One,2013,8(12):e81990.
[10]Wang M,Wang Y,Yan Y,et al.Clinical and laboratory profiles of refractory Mycoplasma pneumoniae pneumonia in children[J].Int J Infect Dis,2014,29:18-23.
[11]Kobayashi Y,Wada H,Rossios C,et al.A novel macrolide solithromycin exerts superior anti-inflammatory effect via NF-κB inhibition[J].J Pharmacol Exp Ther,2013,345(1):76-84.
[12]Saraya T,Nakata K,Nakagaki K,et al.Identification of a mechanism for lung inflammation caused by Mycoplasma pneumoniae using a novel mouse model[J].Results Immunol,2011,1(1):76-87.
[13]Russo RC,Garcia CC,Teixeira MM,et al.The CXCL8/IL-8 chemokine family and its receptors in inflammatory diseases[J].Expert Rev Clin Immunol,2014,10(5):593-619.
[14]毕慧娟,王健,黄河胜,等.慢性乙肝患者外周血单个核细胞中CXCR1、CXCR2及IL-8 mRNA表达与干扰素治疗关系[J].细胞与分子免疫学杂志,2012,28(4):422-425.
[15]Jornot L,Cordey S,Caruso A,et al.T lymphocytes promote the antiviral and inflammatory responses of airway epithelial cells[J].PLo S One,2011,6(10):e26293.
[16]Liu L,Gao Z,Xia C,et al.Comparative study of trans-oral and trans-tracheal intratracheal instillations in a murine model of acute lung injury[J].Anat Rec(Hoboken),2012,295(9):1513-1519.
[17]Jundi K,Greene CM.Transcription of interleukin-8:how altered regulation can affect cystic fibrosis lung disease[J].Biomolecules,2015,5(3):1386-1398.
[18]Gottipati KR,Bandari SK,Nonnenmann MW,et al.Transcriptional mechanisms and protein kinase signaling mediate organic dust induction of IL-8 expression in lung epithelial and THP-1 cells[J].Am J Physiol Lung Cell Mol Physiol,2015,308(1):L11-L21.
[19]Wang J,Lu Q.Expression of T subsets and m IL-2R in peripheral blood of newborns with hypoxic ischemic encephalopathy[J].World J Pediatr,2008,4(2):140-144.
[20]Tsivkovskii R,Sabet M,Tarazi Z,et al.Levofloxacin reduces inflammatory cytokine levels in human bronchial epithelia cells:implications for aerosol MP-376(levofloxacin solution for inhalation)treatment of chronic pulmonary infections[J].FEMS Immunol Med Microbiol,2011,61(2):141-146.
[21]Curatolo W,Foulds G,Labadie R.Mechanistic study of the azithromycin dosage-form-dependent food effect[J].Pharm Res,2010,27(7):1361-1366.
[22]Tanaka R,Sanada S,Fujimura M,et al.Ventilatory impairment detection based on distribution of respiratory-induced changes in pixel values in dynamic chest radiography:a feasibility study[J].Int J Comput Assist Radiol Surg,2011,6(1):103-110.
[23]舒林华,尚云晓,蔡栩栩,等.肺炎支原体肺炎患儿支气管肺泡灌洗液和血清中肺表面活性蛋白的变化及意义[J].中国当代儿科杂志,2012,14(12):928-932.
[24]马香,丁明杰,赵秀侠,等.不同胸部影像学表现的肺炎支原体肺炎儿童的肺功能变化特征[J].中国当代儿科杂志,2014,16(10):997-1000.